Advances in molecular technologies over the past few decades have led to the development of very powerful tools to purify and analyze nucleic acids. Current processes to study RNA transactions such as transcription, translation and processing (e.g., Northern blot, RNA microarray, RNA sequencing and Reverse Transcription RT-PCR analysis) can be complex and time-consuming, and often require specialized instrumentation or trained personnel. These procedures require reagents that have temperature-specific storage requirements and are laboratory-based, which limit their portability and utility for POC testing, especially in resource-poor settings. To facilitate POC analysis, it i critical to integrate sample preparation, sample handling, amplification (PCR; RT-PCR) into a single, automated process. However, few systems have successfully integrated all of these steps. Due to the complexity of integrating sample prep, it has largely been omitted from these systems and left to bench-top equipment. The overall goal of this project is to develop a novel process that can perform a complete, automated, sample-to-answer RNA detection analysis and provide an accurate point-of-care (POC) diagnosis of infection using an RNA virus. During our initial Phase I, feasibility studies will focus on the purification and detection of genomic dengue RNA. We will introduce a novel automated method for purifying viral genomic RNA from multiple organisms (SPM) and analyzing that RNA using low-power RT-PCR techniques in Lynntech's convective amplification unit. This method will allow the diagnosis of the febrile phase of dengue infection and differentiate among serotypes of dengue. The process requires minimal training and provides a sample-to-answer system that simplifies operation, mitigates risk of user error, and decreases overall time-to-detection. Furthermore, the process is not tied to the laboratory, and therefore is conducive to performing RNA studies in-the-field by non-specialized personnel. The proposed process will enable scientists to purify RNA, perform RT and analyze sequences via RT-PCR and will be applicable to transcriptional analysis, RNA detection and POC diagnostics. Aims of the Phase I include: 1) develop a portable RNA sample preparation module; 2) develop a portable RT-PCR assay to identify RNA using convective flow amplification; and 3) automate the sample prep device. Phase II will focus on adapting the sample-to-answer RNA detection process to a real-time POC detection system. We will integrate SPM and convective amplification systems into a single portable unit that can be used beyond the confines of the traditional laboratory to study RNA processes, as well as to diagnose infections such as dengue. Lynntech's real-time detection system will be a next generation tool for delivering molecular diagnostics into the hands of health-care professionals globally. This system will provide necessary resources to communities and countries where traditional tests are not available. The final product is envisaged to be an FDA-approved, CLIA-waived RNA-based sample-to-answer diagnostic device based on the process established in Phase I and II.